Functional properties of insect olfactory receptors: ionotropic receptors and odorant receptors

The majority of insect olfactory receptors belong to two distinct protein families, the ionotropic receptors (IRs), which are related to the ionotropic glutamate receptor family, and the odorant receptors (ORs), which evolved from the gustatory receptor family. Both receptor types assemble to heteromeric ligand-gated cation channels composed of odor-specific receptor proteins and co-receptor proteins. We here present in short the current view on evolution, function, and regulation of IRs and ORs. Special attention is given on how their functional properties can meet the environmental and ecological challenges an insect has to face

Homeostasis of mitochondrial Ca²⁺ stores is critical for signal amplification in Drosophila melanogaster olfactory sensory neurons

SIMPLE SUMMARY: The evolution of flight imposed new challenges on insects when locating and identifying food sources, mates, or enemies. As an adaptation, flying insects developed a remarkably sensitive olfactory system to detect faint odor traces. This ability is linked to the olfactory receptor class of odorant receptors, which are found in insect olfactory sensory neurons. In a subgroup of these neurons, sensitivity can be further enhanced through a process called sensitization. Extracellular calcium ions, calmodulin, and protein kinase C are known to be key factors in this process. While manipulation of mitochondrial calcium im- and export has been shown to influence odor responses in general, the connection of intracellular calcium stores to sensitization has so far been only speculative. Using two pharmacological approaches, we disrupted mitochondrial calcium management in order to explore its importance to sensitization. Overall, our findings reveal that mitochondrial calcium stores are important players in the complex intracellular signaling pathways required for sensitization. ABSTRACT: Insects detect volatile chemosignals with olfactory sensory neurons (OSNs) that express olfactory receptors. Among them, the most sensitive receptors are the odorant receptors (ORs), which form cation channels passing Ca(2+). OSNs expressing different groups of ORs show varying optimal odor concentration ranges according to environmental needs. Certain types of OSNs, usually attuned to high odor concentrations, allow for the detection of even low signals through the process of sensitization. By increasing the sensitivity of OSNs upon repetitive subthreshold odor stimulation, Drosophila melanogaster can detect even faint and turbulent odor traces during flight. While the influx of extracellular Ca(2+) has been previously shown to be a cue for sensitization, our study investigates the importance of intracellular Ca(2+) management. Using an open antenna preparation that allows observation and pharmacological manipulation of OSNs, we performed Ca(2+) imaging to determine the role of Ca(2+) storage in mitochondria. By disturbing the mitochondrial resting potential and induction of the mitochondrial permeability transition pore (mPTP), we show that effective storage of Ca(2+) in the mitochondria is vital for sensitization to occur, and release of Ca(2+) from the mitochondria to the cytoplasm promptly abolishes sensitization. Our study shows the importance of cellular Ca(2+) management for sensitization in an effort to better understand the underlying mechanics of OSN modulation

Beyond Pearson’s correlation: modern nonparametric independence tests for psychological research

When examining whether two continuous variables are associated, tests based on Pearson’s, Kendall’s, and Spearman’s correlation coefficients are typically used. This paper explores modern nonparametric independence tests as an alternative, which, unlike traditional tests, have the ability to potentially detect any type of relationship. In addition to existing modern nonparametric independence tests, we developed and considered two novel variants of existing tests, most notably the Heller-Heller-Gorfine-Pearson (HHG-Pearson) test. We conducted a simulation study to compare traditional independence tests, such as Pearson’s correlation, and the modern nonparametric independence tests in situations commonly encountered in psychological research. As expected, no test had the highest power across all relationships. However, the distance correlation and the HHG-Pearson tests were found to have substantially greater power than all traditional tests for many relationships and only slightly less power in the worst case. A similar pattern was found in favor of the HHG-Pearson test compared to the distance correlation test. However, given that distance correlation performed better for linear relationships and is more widely accepted, we suggest considering its use in place or additional to traditional methods when there is no prior knowledge of the relationship type, as is often the case in psychological research

A computer simulation-based analysis of supply chains resilience in industrial environment

The article presents a computer simulation-based model for analysis of supply chain resilience, which allows determining and verifying the generally valid principles, capabilities and ways for building long-term resilience of global supply chains against serious disruptions. The model is created on the basis of a supply chain from automotive industry and contains the main logistics flows used by present automotive producers. Any real automotive supply chain can be modelled as a combination of these logistics flows. The model was created in DOSIMIS-3® and verified using experimental data. The performed simulation demonstrates a significant decrease of the supply chain performance in case of serious disruption occurrence

Calmodulin affects sensitization of Drosophila melanogaster odorant receptors

Flying insects have developed a remarkably sensitive olfactory system to detect faint and turbulent odor traces. This ability is linked to the olfactory receptors class of odorant receptors (ORs), occurring exclusively in winged insects. ORs form heteromeric complexes of an odorant specific receptor protein (OrX) and a highly conserved co-receptor protein (Orco). The ORs form ligand gated ion channels that are tuned by intracellular signaling systems. Repetitive subthreshold odor stimulation of olfactory sensory neurons sensitizes insect ORs. This OR sensitization process requires Orco activity. In the present study we first asked whether OR sensitization can be monitored with heterologously expressed OR proteins. Using electrophysiological and calcium imaging methods we demonstrate that D. melanogaster OR proteins expressed in CHO cells show sensitization upon repeated weak stimulation. This was found for OR channels formed by Orco as well as by Or22a or Or56a and Orco. Moreover, we show that inhibition of calmodulin (CaM) action on OR proteins, expressed in CHO cells, abolishes any sensitization. Finally, we investigated the sensitization phenomenon using an ex vivo preparation of olfactory sensory neurons (OSNs) expressing Or22a inside the fly's antenna. Using calcium imaging, we observed sensitization in the dendrites as well as in the soma. Inhibition of calmodulin with W7 disrupted the sensitization within the outer dendritic shaft, whereas the sensitization remained in the other OSN compartments. Taken together, our results suggest that CaM action is involved in sensitizing the OR complex and that this mechanisms accounts for the sensitization in the outer dendrites, whereas further mechanisms contribute to the sensitization observed in the other OSN compartments. The use of heterologously expressed OR proteins appears to be suitable for further investigations on the mechanistic basis of OR sensitization, while investigations on native neurons are required to study the presently unknown additional mechanisms involved in OSN sensitization

Functional interaction between Drosophila olfactory sensory neurons and their support cells

Insects detect volatile chemicals using antennae, which house a vast variety of olfactory sensory neurons (OSNs) that innervate hair-like structures called sensilla where odor detection takes place. In addition to OSNs, the antenna also hosts various support cell types. These include the triad of trichogen, tormogen, and thecogen support cells that lie adjacent to their respective OSNs. The arrangement of OSN supporting cells occurs stereotypically for all sensilla and is widely conserved in evolution. While insect chemosensory neurons have received considerable attention, little is known about the functional significance of the cells that support them. For instance, it remains unknown whether support cells play an active role in odor detection, or only passively contribute to homeostasis, e.g., by maintaining sensillum lymph composition. To investigate the functional interaction between OSNs and support cells, we used optical and electrophysiological approaches in Drosophila. First, we characterized the distribution of various supporting cells using genetic markers. By means of an ex vivo antennal preparation and genetically-encoded Ca(2+) and K(+) indicators, we then studied the activation of these auxiliary cells during odor presentation in adult flies. We observed acute responses and distinct differences in Ca(2+) and K(+) fluxes between support cell types. Finally, we observed alterations in OSN responses upon thecogen cell ablation in mature adults. Upon inducible ablation of thecogen cells, we notice a gain in mechanical responsiveness to mechanical stimulations during single-sensillum recording, but a lack of change to the neuronal resting activity. Taken together, these results demonstrate that support cells play a more active and responsive role during odor processing than previously thought. Our observations thus reveal that support cells functionally interact with OSNs and may be important for the extraordinary ability of insect olfactory systems to dynamically and sensitively discriminate between odors in the turbulent sensory landscape of insect flight